Anti-UBE3A Picoband™ Antibody (monoclonal, 3F13)

Product Name

Anti-UBE3A Picoband™ Antibody (monoclonal, 3F13)

View all E6AP/UBE3A Antibodies

SKU/Catalog Number

M00582-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-UBE3A Picoband™ Antibody (monoclonal, 3F13) catalog # M00582-1. Tested in Flow Cytometry, WB applications. This antibody reacts with Human, Monkey.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-UBE3A Picoband™ Antibody (monoclonal, 3F13) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00582-1)

Host

Mouse

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

Clonality

Monoclonal

Clone Number

3F13

Isotype

Mouse IgG2b

Immunogen

E.coli-derived human UBE3A recombinant protein (Position: M1-E860).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M00582-1 is reactive to UBE3A in Human, Monkey

Applications

M00582-1 is guaranteed for Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

100 kDa

Calculated molecular weight

100.688kDa

Background of E6AP/UBE3A

Ubiquitin-protein ligase E3A (UBE3A) also known as E6AP ubiquitin-protein ligase (E6AP) is an enzyme that in humans is encoded by the UBE3A gene. It is mapped to 15q11.2. This gene encodes an E3 ubiquitin-protein ligase, part of the ubiquitin protein degradation system. This imprinted gene is maternally expressed in brain and biallelically expressed in other tissues. Maternally inherited deletion of this gene causes Angelman Syndrome, characterized by severe motor and intellectual retardation, ataxia, hypotonia, epilepsy, absence of speech, and characteristic facies. The protein also interacts with the E6 protein of human papillomavirus types 16 and 18, resulting in ubiquitination and proteolysis of tumor protein p53. Alternative splicing of this gene results in three transcript variants encoding three isoforms with different N-termini. Additional transcript variants have been described, but their full length nature has not been determined.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Human, Monkey
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of UBE3A using anti-UBE3A antibody (M00582-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human HEK293 whole cell lysates,
Lane 3: human Raji whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-UBE3A antigen affinity purified monoclonal antibody (Catalog # M00582-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for UBE3A at approximately 100KD. The expected band size for UBE3A is at 100KD.

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Figure 2. Flow Cytometry analysis of U87 cells using anti-UBE3A antibody (M00582-1).
Overlay histogram showing U87 cells stained with M00582-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-UBE3A Antibody (M00582-1, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For UBE3A (Source: Uniprot.org, NCBI)

Alternative Names

ANCR; ANCREPVE6AP; AS; CTCL tumor antigen se37-2; E6AP ubiquitin-protein ligase; E6AP; E6-AP; EC 6.3.2.-; EPVE6AP; FLJ26981; HPVE6A; human papilloma virus E6-associated protein; Human papillomavirus E6-associated protein; Oncogenic protein-associated protein E6-AP; Renal carcinoma antigen NY-REN-54; UBE3A; ubiquitin protein ligase E3A; ubiquitin-protein ligase E3A UBE3A ANCR, AS, E6-AP, EPVE6AP, HPVE6A ubiquitin protein ligase E3A ubiquitin-protein ligase E3A|CTCL tumor antigen se37-2|E6AP ubiquitin-protein ligase|HECT-type ubiquitin transferase E3A|human papilloma virus E6-associated protein|human papillomavirus E6-associated protein|oncogenic protein-associated protein E6-AP|renal carcinoma antigen NY-REN-54

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In this infographic, you will see the following information for UBE3A: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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